Effects of cryopreservation on the immunogenicity of porcine arterial allografts in early stages of transplant vasculopathy

doi:10.1016/j.cryobiol.2005.06.006

Cryobiology

Volume 51, Issue 2, October 2005, Pages 130-141

https://doi.org/10.1016/j.cryobiol.2005.06.006 Get rights and content

Abstract

Background

The number of revascularization procedures including coronary and lower extremity bypass, have increased greatly in the last decade. It suggests a growing need for vascular grafts. Cryopreserved allografts could represent a viable alternative but their immunologic reactivity remains controversial.

Methods

71 pigs (40 recipients and 31 donors) were used. Two femoral grafts per recipient animal were implanted for 3, 7, and 30 days. Types of grafts: fresh autograft as a control graft (n = 19), fresh allograft (n = 31) and cryopreserved allograft (n = 30). Histological and immunohistochemical studies were performed.

Results

Fresh allografts compared to autografts showed intimal inflammatory infiltration at 3 days (328 vs. 0 macrophages/mm²; P < 0.05) and 7 days (962 vs. 139 T lymphocytes/mm²; P < 0.05) post-transplantation. At 30 days, there was a loss of endothelial cells, presence of luminal thrombus and aneurismal lesions (total area = 15.8 vs. 8.4 mm²; P < 0.05). Cryopreservation did not reduce these lesions nor modify endothelial nitric oxide synthase (eNOS) expression nor modify the number of animals that developed anti-SLA antibodies. Moreover, at 7 days, cryopreserved allografts compared to fresh allografts showed a higher expression of P-selectin (5 out of 5 vs. 1 out of 5; P < 0.05) and, at 30 days, a greater inflammatory reactivity (2692 vs. 1107 T lymphocytes/mm² in media; P < 0.05) with a trend towards a higher presence of multinucleated giant cells than in the fresh ones.

Conclusions

The cryopreservation method used maintained immunogenicity of allografts and increased the inflammatory reactivity found in fresh allografts up to 30 days of vascular transplantation.

Section snippets

Animals

Forty-nine Landrace × Large White pigs (29 recipients and 20 donors) were used for the main part of the study and a set of 22 additional pigs (11 recipients and 11 donors) were used to determine the differences in eNOS and P-selectin (CD62P) expression and alloantibody formation between fresh and cryopreserved allografts. The study was approved by the Institutional Review Board and Ethics Committee of our institution; animal care complied with the Guide for the Care and Use of Laboratory Animals

Results

One of the autografts implanted for 30 days and one of the cryopreserved allografts implanted for 7 days from the first set of animals were found occluded due to surgically related problems and they were excluded from the histological study. All other grafts from both sets of animals were patent at the end of follow-up.

Discussion

The results of this study demonstrate that, in our model, the persistent presence of inflammatory cells due to transplant vasculopathy found in fresh allografts produced endothelial cells loss, luminal thrombus, and aneurismal lesions. On the other hand, cryopreservation neither reduced these lesions nor eNOS expression nor the number of animals that developed anti-SLA antibodies. Moreover, cryopreserved allografts showed a higher expression of P-selectin and a greater inflammatory reactivity

Acknowledgments

We thank Josep Maria Manresa of the IMIM for his advice on statistical analysis and Montse Vázquez and Cristina Siles from Hospital Clínic for secretarial assistance. We thank Isabel Rojo for assistance with Flow Cytometry Crossmatch.

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During lobectomy, resection of pulmonary artery, followed by reconstruction or replacement with or without concomitant sleeve bronchial resection, is feasible in selected cases. We report morbidity, mortality, and technical issues in pulmonary artery replacement using a cryopreserved arterial allograft after sleeve resection for centrally located non–small cell lung carcinoma (NSCLC).
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^☆: Statement of funding: This work was financed in part by a grant from Fundació la Marató de TV3 (004210, 2000) by a grant from Sociedad Española de Cardiologia (2002–2004) and by Ministerio de Sanidad y Consumo, Instituto de Salud Carlos III, Red RECAVA.

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Effects of cryopreservation on the immunogenicity of porcine arterial allografts in early stages of transplant vasculopathy☆

Abstract

Background

Methods

Results

Conclusions

Section snippets

Animals

Results

Discussion

Acknowledgments

J. Thorac. Cardiovasc. Surg.

Cryobiology

J. Vasc. Surg.

Cryobiology

J. Vasc. Surg.

Eur. J. Vasc. Endovasc. Surg.

Ann. Vasc. Surg.

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Vet. Immunol. Immunopathol.

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